CN112776325B - Three-dimensional ultrasonic array support-free cell printing device and printing process thereof - Google Patents
Three-dimensional ultrasonic array support-free cell printing device and printing process thereof Download PDFInfo
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- CN112776325B CN112776325B CN202011601163.7A CN202011601163A CN112776325B CN 112776325 B CN112776325 B CN 112776325B CN 202011601163 A CN202011601163 A CN 202011601163A CN 112776325 B CN112776325 B CN 112776325B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
- B29C64/20—Apparatus for additive manufacturing; Details thereof or accessories therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
- B29C64/30—Auxiliary operations or equipment
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
- B29C64/30—Auxiliary operations or equipment
- B29C64/307—Handling of material to be used in additive manufacturing
- B29C64/321—Feeding
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
- B29C64/30—Auxiliary operations or equipment
- B29C64/35—Cleaning
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y30/00—Apparatus for additive manufacturing; Details thereof or accessories therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y40/00—Auxiliary operations or equipment, e.g. for material handling
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
Abstract
The three-dimensional ultrasonic array cell printing device without the support and the printing process thereof comprise a cell conveying mechanism, a printing platform, a platform driving mechanism, a conveying driving mechanism and a three-dimensional ultrasonic array micro-potential well generating system, wherein the printing platform is installed on the platform driving mechanism, the cell conveying mechanism is installed on the conveying driving mechanism, the three-dimensional ultrasonic array micro-potential well generating system is connected with the printing platform, a planar sound pressure micro-potential well is formed in the printing platform, a plurality of planar sound pressure micro-potential wells with different heights are formed on the printing platform, so that cells needing to be printed at different heights are positioned, a cell three-dimensional system is constructed by adopting a mode of constructing the three-dimensional array sound pressure potential well, the three-dimensional cell printing device is better in controllability, can be used for three-dimensional printing of cells under various environments, and has excellent universality.
Description
Technical Field
The invention belongs to the fields of biological manufacturing and cell printing, and particularly relates to a three-dimensional ultrasonic array stentless cell printing device and a printing process thereof.
Background
The cell/biological 3D printing is to carry out three-dimensional assembly of cells by gradually depositing living cells, extracellular matrix, biological factors and necessary biological materials, construct a three-dimensional multi-cell system with biological activity in vitro, and then achieve the purposes of tissue regeneration and organ repair through culture and propagation, thereby being an effective way for solving the dilemma of the traditional tissue engineering and having great significance for the development of life science and biological medicine.
In the cell printing technology, the problem to be solved is how to accurately deposit a set number of specific cells to a preset position in a three-dimensional space, i.e., how to construct a three-dimensional system of cells as required, and the method is favorable for nutrient delivery of cell culture and propagation, and finally forms tissues. In order to achieve the above purpose, in the conventional cell 3D printing, a scaffold material represented by hydrogel is usually mixed with a cell sap to prepare a bio-ink, and the bio-ink is printed, and the hydrogel is solidified to form a cell-attached biological scaffold. However, as a carrier for further propagation of cells, the biological scaffold has many challenges, such as interference of cell-cell communication, non-uniform cell attachment, influence of scaffold degradation, and poor precision of scaffold materials. The existing scaffold-free cell printing technology avoids doping a scaffold material into cell sap to prepare biological ink for printing, and has obvious advantages in the aspects of cell-cell communication and the like. However, the existing stentless cell printing technology still needs to use agar rods and the like as physical supports in the cell printing process. Therefore, the research on the cell 3D printing technology without a bracket and a support is of great significance.
Disclosure of Invention
Aiming at the defects, the technical problem to be solved by the invention is to provide a three-dimensional ultrasonic array stentless cell printing device and a printing process thereof, wherein a three-dimensional multi-cell system with bioactivity is constructed in vitro by gradually depositing living cells, extracellular matrix, biological factors and necessary biological materials to carry out three-dimensional assembly of the cells, and then the purposes of tissue regeneration and organ repair are achieved through culture and propagation.
In order to solve the technical problems, the invention adopts the technical scheme that,
the three-dimensional ultrasonic array stentless cell printing device comprises a cell conveying mechanism, a printing platform, a platform driving mechanism, a conveying driving mechanism and a three-dimensional ultrasonic array micro-potential well generating system, wherein the printing platform is installed on the platform driving mechanism, the cell conveying mechanism is installed on the conveying driving mechanism, the three-dimensional ultrasonic array micro-potential well generating system is connected with the printing platform, and a planar sound pressure micro-potential well array is formed in the printing platform.
Furthermore, the three-dimensional ultrasonic array micro-potential well generating system comprises a signal generator, a power amplifier and a piezoelectric transducer array, wherein the piezoelectric transducer array is fixedly installed on the printing platform, the power amplifier is connected with the piezoelectric transducer array, and the signal generator is connected with the power amplifier.
Furthermore, the printing platform comprises a chassis and a polygonal resonant cavity, the chassis is connected with the platform driving mechanism, the polygonal resonant cavity is fixedly installed on the chassis, the piezoelectric transducer array is fixedly installed on the inner wall of the polygonal resonant cavity, and a planar sound pressure micro-potential well array is formed in the polygonal resonant cavity.
Furthermore, a piezoelectric transducer array is arranged on the inner wall of the polygonal resonant cavity and arranged at equal intervals in the height direction to form a plurality of sound pressure micro-potential wells with different heights.
Furthermore, the cell conveying mechanism comprises a micro-conveying nozzle, a pulse actuating device, a nozzle clamp and a frame plate, wherein the micro-conveying nozzle is fixedly arranged on the frame plate through the nozzle clamp, and the pulse actuating device is fixedly arranged on the frame plate and is connected with the upper part of the micro-conveying nozzle.
Furthermore, a spray head is installed at the lower end of the micro-delivery nozzle, and a cell counter is fixedly installed between the micro-delivery nozzle and the spray head.
Furthermore, a microscope is fixedly mounted on the frame plate and arranged towards the spray head.
Further, platform actuating mechanism includes bed plate, lifting support, lift lead screw sliding block set spare and elevator motor, and lifting support fixed mounting is on the bed plate, and lift lead screw sliding block set spare rotates to be installed on lifting support, and elevator motor is connected with lift lead screw sliding block set spare.
Further, carry actuating mechanism to include X axle driving piece, Y axle driving piece and support column, support column fixed mounting is on the bed plate, Y axle driving piece fixed mounting is on the support column, X axle driving piece fixed mounting is on Y axle driving piece, and cell conveying mechanism installs on X axle driving piece.
The printing process of the three-dimensional ultrasonic array stent-free cell printing device comprises the following steps,
s1: sterilizing the printing device, keeping the temperature constant and proper, injecting culture solution into the polygonal resonant cavity, and injecting cell suspension into the micro-nozzle;
s2: according to a pre-designed model, the piezoelectric transducer array works, and a planar array sound pressure micro-potential well is excited on the lowest layer of the polygonal resonant cavity as required;
s3: according to a pre-designed model, the platform driving mechanism is controlled to move, so that the spray head reaches a specific height required by printing of the polygonal resonant cavity model, and then the conveying driving mechanism is controlled to move, so that the micro-nozzle moves to a specified sound pressure micro-potential well position;
s4: according to the requirement of a model, a cell conveying mechanism conveys a specific number of cells to the corresponding planar array sound pressure micro-potential well, the conveyed cells are captured in the sound pressure potential well under the action of the sound restoring force in the sound pressure potential well, the printing of the cells in the single potential well is completed, and the cell printing in all the sound potential wells in one layer of plane is completed by sub-class pushing;
s5: after the first layer is completely printed, the three-dimensional ultrasonic array micro-potential well generating system excites the array sound pressure micro-potential well of the second layer, the polygonal culture platform is moved downwards to continue printing, the steps S2 to S4 are repeated, and the printing of the cell three-dimensional system is realized by analogy;
s6: and after printing, carrying out cell culture on the cell three-dimensional system at a proper temperature, and finally forming cell tissues through cell division and growth.
The beneficial effect of the invention is that,
the invention realizes the construction of a three-dimensional system of cells by controlling the cells by using the acoustic radiation force, and the acoustic radiation force control cells have remarkable advantages in the aspects of universality, biocompatibility, control scale and the like. The cell three-dimensional control system has no special requirements on cell types, density, shapes and the like, can be spread in a light-tight medium, has stronger applicability, and can complete three-dimensional control of various cells under various environments. The method adopts a mode of constructing the three-dimensional array sound pressure potential well to construct the cell three-dimensional system, has better controllability, can be used for three-dimensional printing of cells under various cells and various environments, and has excellent universality.
The invention constructs a three-dimensional array sound pressure micro-potential well through an array transducer, captures cell particles by using sound radiation force, and the micro-potential well is a cell capture point (node), wherein the node is arranged into a line, the line is arranged into a surface, and the surface is overlapped to form a body, thereby constructing a cell three-dimensional system and forming cell tissues through culture. The technology does not need biological scaffolds/supports, and has remarkable advantages in the aspects of cell density, intercellular communication, cell growth and the like. And the scaffold/support degradation process is avoided, degradation byproducts are avoided, and cells are more easily survived and grown. The cell printing process does not generate the phenomena of high temperature, high pressure, strong electric field and the like, the survival rate of the printed cells is further improved, and the survival rate can be ensured to be more than 90 percent.
The technology for driving the cell printing by the pulse inertia force is adopted, the technology can realize the accurate conveying of a single cell or a plurality of cells by changing the size of a single pulse through the pulse inertia force generated by the pulse actuating device, realize the accurate and controllable conveying of the cells in the printing process, ensure the efficient and accurate construction of a three-dimensional system of the cells, and has important in-vitro bioengineering application potential.
Drawings
Fig. 1 is a schematic diagram of a three-dimensional ultrasound array micro-well stentless cell 3D printing system.
FIG. 2 is a partially enlarged schematic view of a polygonal resonator.
Fig. 3 to 5 are schematic diagrams of single-cell and multi-cell printing in a single acoustic potential well.
Fig. 6 to 9 are schematic diagrams of the three-dimensional ultrasonic array micro-potential well stent-free cell 3D printing process.
Description of the drawings: 1. a signal generator; 2. a power amplifier; 3. an observation microscope; 4. a micro-delivery nozzle; 5. a pulse actuating device; 6. a nozzle holder; 7. a frame plate; 8. a chassis; 9. an ultraviolet sterilizing lamp; 12 a cell counter; 13 an array of piezoelectric transducers; 14. a polygonal resonant cavity; 15. an upper computer; 16. and (4) a spray head.
Detailed Description
The invention is further described below with reference to the accompanying drawings.
Three-dimensional ultrasonic array does not have support cell printing device, including cell conveying mechanism A, print platform B, platform actuating mechanism C, carry actuating mechanism D and three-dimensional ultrasonic array micro-potential well generation system E, print platform B installs on platform actuating mechanism C, cell conveying mechanism A installs on carrying actuating mechanism D, three-dimensional ultrasonic array micro-potential well generation system E is connected with print platform B, form plane sound pressure micro-potential well array in print platform B, form a plurality of plane sound pressure micro-potential well arrays of different height on print platform B, it fixes a position to realize the cell that needs to print at different heights, adopt the mode of constructing three-dimensional array sound pressure potential well to construct three-dimensional system of cell, controllability is better, can be used for multiple cell, cell three-dimensional printing under the multiple environment, have fabulous universality.
The three-dimensional ultrasonic array micro-potential well generating system comprises a signal generator 1, a power amplifier 2 and a piezoelectric transducer array 13, wherein the piezoelectric transducer array 13 is fixedly arranged on a printing platform B, the power amplifier 2 is connected with the piezoelectric transducer array 13, the signal generator 1 is connected with the power amplifier 2, the piezoelectric transducer array is fixed on the inner wall of a culture cavity and is arranged at equal intervals in the height direction, and a plurality of transducers of each layer are on the same horizontal plane, so that a plurality of planar sound pressure micro-potential well arrays with different heights are formed; the front end of the signal generator 2 is connected with an upper computer 15, the signal generator 2 generates signals with specific frequency and specific waveform under the control of the upper computer 15, and the wave signals are amplified into electric signals with specific voltage through a power amplifier according to requirements and then transmitted to a piezoelectric transducer array of a layer of the array piezoelectric transducer array; the piezoelectric transducer array is driven by the electric signal to generate sound waves with specific energy and specific frequency to form a polygonal plane sound field and construct a plane sound pressure micro-potential well array; the array piezoelectric transducer array forms different planar array sound pressure micro-potential wells by exciting piezoelectric transducer arrays on different layers, and the planar array sound pressure micro-potential wells are built and superposed layer by layer to form a three-dimensional array sound pressure micro-potential well.
The printing platform comprises a chassis 8 and a polygonal resonant cavity 14, the chassis 8 is connected with a platform driving mechanism C, the polygonal resonant cavity 14 is fixedly mounted on the chassis 8, a piezoelectric transducer array 13 is fixedly mounted on the inner wall of the polygonal resonant cavity 14, a planar sound pressure micro-potential well array is formed in the polygonal resonant cavity 14, transducers with different heights are excited to form a plurality of planar array sound pressure potential wells, and the planar array sound pressure potential wells are superposed to form a three-dimensional array sound pressure potential well.
The piezoelectric transducer array 13 is installed on the inner wall of the polygonal resonant cavity 14, and the piezoelectric transducer array 13 is arranged in the height direction at equal intervals to form a plurality of sound pressure micro-potential wells with different heights, and the sound pressure micro-potential wells are superposed to form a three-dimensional array sound pressure potential well.
Cell conveying mechanism A is including carrying nozzle 4 a little, pulse actuating device 5, nozzle holder 6, frame plate 7, it passes through nozzle holder 6 fixed mounting on frame plate 7 to carry nozzle 4 a little, pulse actuating device 5 fixed mounting is on frame plate 7, and be connected on 4 upper portions of carrying nozzle a little, pulse inertial force through the pulse actuating device production, size through changing single pulse, can realize the accurate transport of single cell or a plurality of cells, it is accurate controllable to realize printing in-process cell transport, the high-efficient accurate of having guaranteed the three-dimensional system of cell constructs.
A nozzle 16 is installed at the lower end of the micro-delivery nozzle 4, a cell counter 12 is fixedly installed between the micro-delivery nozzle 4 and the nozzle 16, and the delivery is stopped when the number of delivered cells reaches a predetermined number.
Fixed mounting has microscope 3 on the frame plate 7, and microscope 3 sets up towards shower nozzle 16, and the shower nozzle motion is followed to the microscope for observe cell position state when printing, microscope 3 is connected with host computer 15, host computer 15 is according to the transport of the motion of microscope 3's detection feedback control three-dimensional platform and cell.
The platform driving mechanism C comprises a base plate 1-1, a lifting support 1-2, a lifting lead screw sliding block assembly 1-3 and a lifting motor 1-4, wherein the lifting support 1-2 is fixedly installed on the base plate 1-1, the lifting lead screw sliding block assembly 1-3 is rotatably installed on the lifting support 1-2, the lifting motor 1-4 is connected with the lifting lead screw sliding block assembly 1-3, the printing platform B is fixedly installed on a sliding block of the lifting lead screw sliding block assembly 1-3, and the printing platform B is subjected to height adjustment through the platform driving mechanism C.
The conveying driving mechanism D comprises an X-axis driving piece, a Y-axis driving piece and a supporting column 2-1, the supporting column 2-1 is fixedly installed on the base plate 1-1, the Y-axis driving piece is fixedly installed on the supporting column 1-1, the X-axis driving piece is fixedly installed on the Y-axis driving piece, the cell conveying mechanism is installed on the X-axis driving piece, and the plane position of the cell conveying mechanism A is adjusted through the X-axis driving piece and the Y-axis driving piece.
In some preferred modes, the structures of the X-axis driving part and the Y-axis driving part are consistent with the structure of the platform driving mechanism C, and both comprise a bracket, a lead screw slider assembly and a driving motor, the lead screw slider assembly is rotatably installed in the bracket, and the driving motor is connected with the lead screw slider assembly.
In some preferred modes, a cover is arranged on the device, and an ultraviolet sterilizing lamp 9 is fixedly arranged on the cover.
The printing process of the three-dimensional ultrasonic array stentless cell printing device comprises the following steps,
s1: sterilizing the printing device, keeping the temperature constant and proper, injecting culture solution into the polygonal resonant cavity, and injecting cell suspension into the micro-nozzle;
s2: according to a pre-designed model, the piezoelectric transducer array works, and a planar array sound pressure micro-potential well is excited on the lowest layer of the polygonal resonant cavity as required;
s3: according to a pre-designed model, the driving mechanism of the control platform moves to enable the spray head to reach a specific height required by printing of the polygonal resonant cavity model, and then the conveying driving mechanism is controlled to move to enable the micro-nozzle to move to a specified sound pressure micro-potential well position;
s4: according to the requirement of a model, a cell conveying mechanism conveys a specific number of cells to the corresponding planar array sound pressure micro-potential well, the conveyed cells are captured in the sound pressure potential well under the action of the sound restoring force in the sound pressure potential well, the printing of the cells in the single potential well is completed, and the cell printing in all the sound potential wells in one layer of plane is completed by sub-class pushing;
s5: after the first layer is completely printed, the three-dimensional ultrasonic array micro-potential well generating system excites the array sound pressure micro-potential well of the second layer, the polygonal culture platform is moved downwards to continue printing, the steps S2 to S4 are repeated, and the printing of the cell three-dimensional system is realized by analogy;
s6: and after printing, performing cell culture on the cell three-dimensional system at a proper temperature, and finally forming cell tissues through cell division and growth.
When cell printing is carried out, the cell conveying mechanism A outputs a single or a plurality of printing cells, the cells can be printed according to model requirements, the cell liquid concentration is higher when the cells are printed, and the cell liquid concentration loaded in the micro-nozzle is about 10 6 ～10 7 And adjusting the concentration of the cell sap according to the size of the printed cells.
In the printing process, when the nozzle 16 moves to the designated position of the culture cavity, the nozzle 16 is not suspended above the array acoustic potential well, but is positioned in the potential well, and the micro-nozzle directly prints cells to the proper position in the specific potential well.
In the invention, the array ultrasonic transducer is arranged on the inner wall of the polygonal resonant cavity, can be two surfaces or a plurality of surfaces, when the array ultrasonic transducer is arranged on the plurality of surfaces, the array ultrasonic transducer forms different array sound pressure potential wells through the combination of the transducers on different surfaces, the array ultrasonic transducer can form sound potential wells with different sizes through exciting sound waves with different frequencies, so as to construct array sound pressure micro potential wells with different densities, the exciting frequency of the array ultrasonic transducer is in MHz level, and the size of the formed potential well is in micron level.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention; thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Although the terms corresponding to the reference numerals in the figures are used more herein, the possibility of using other terms is not excluded; these terms are used merely to more conveniently describe and explain the nature of the present invention; they are to be construed as being without limitation to any additional limitations that may be imposed by the spirit of the present invention.
Claims (9)
1. The three-dimensional ultrasonic array stentless cell printing device is characterized by comprising a cell conveying mechanism (A), a printing platform (B), a platform driving mechanism (C), a conveying driving mechanism (D) and a three-dimensional ultrasonic array micro-potential well generating system (E), wherein the printing platform (B) is arranged on the platform driving mechanism (C), the cell conveying mechanism (A) is arranged on the conveying driving mechanism (D), the three-dimensional ultrasonic array micro-potential well generating system (E) is connected with the printing platform (B), and a planar sound pressure micro-potential well array is formed in the printing platform (B);
the three-dimensional ultrasonic array micro-potential well generating system (E) comprises a signal generator (1), a power amplifier (2) and a piezoelectric transducer array (13), wherein the piezoelectric transducer array (13) is fixedly arranged on a printing platform (B), the power amplifier (2) is connected with the piezoelectric transducer array (13), and the signal generator (1) is connected with the power amplifier (2); the front end of the signal generator is connected with an upper computer, the signal generator generates signals with specific frequency and specific waveform under the control of the upper computer, and the wave signals are amplified into electric signals with specific voltage through a power amplifier according to requirements and then transmitted to an array of a certain layer of the piezoelectric transducer array; the piezoelectric transducer array is driven by the electric signal to generate sound waves with specific energy and specific frequency to form a polygonal plane sound field and construct a plane sound pressure micro-potential well array; the piezoelectric transducer array forms different planar array sound pressure micro-potential wells by exciting arrays in different layers, and the planar array sound pressure micro-potential wells are built and superposed layer by layer to form a three-dimensional array sound pressure micro-potential well.
2. The three-dimensional ultrasonic array stent-free cell printing device according to claim 1, wherein the printing platform (B) comprises a base plate (8) and a polygonal resonant cavity (14), the base plate (8) is connected with the platform driving mechanism (C), the polygonal resonant cavity (14) is fixedly installed on the base plate (8), the piezoelectric transducer array (13) is fixedly installed on the inner wall of the polygonal resonant cavity (14), and a planar sound pressure micro-potential well array is formed in the polygonal resonant cavity (14).
3. The three-dimensional ultrasound array stentless cell printing apparatus of claim 2, wherein the piezoelectric transducer arrays (13) are arranged equidistantly in a height direction to form a plurality of sound pressure micro-potential well arrays of different heights.
4. The three-dimensional ultrasonic array stentless cell printing device of claim 1, wherein the cell conveying mechanism (a) comprises a micro-conveying nozzle (4), a pulse actuating device (5), a nozzle clamp (6) and a frame plate (7), the micro-conveying nozzle (4) is fixedly arranged on the frame plate (7) through the nozzle clamp (6), and the pulse actuating device (5) is fixedly arranged on the frame plate (7) and is connected with the upper part of the micro-conveying nozzle (4).
5. The three-dimensional ultrasonic array stentless cell printing device of claim 4, wherein a nozzle (16) is mounted at the lower end of the micro-delivery nozzle (4), and a cell counter (12) is fixedly mounted between the micro-delivery nozzle (4) and the nozzle (16).
6. The three-dimensional ultrasonic array stentless cell printing device of claim 5, wherein the microscope (3) is fixedly mounted on the frame plate (7), the microscope (3) being disposed towards the nozzle (16).
7. The three-dimensional ultrasonic array stentless cell printing apparatus of claim 1, wherein the platform driving mechanism (C) comprises a base plate, a lifting bracket, a lifting lead screw slider assembly and a lifting motor, wherein the lifting bracket is fixedly mounted on the base plate, the lifting lead screw slider assembly is rotatably mounted on the lifting bracket, and the lifting motor is connected with the lifting lead screw slider assembly.
8. The three-dimensional ultrasonic array stentless cell printing apparatus of claim 1, wherein the transport driving mechanism (D) comprises an X-axis driving member, a Y-axis driving member and a support column, the support column is fixedly mounted on the base plate, the Y-axis driving member is fixedly mounted on the support column, the X-axis driving member is fixedly mounted on the Y-axis driving member, and the cell transport mechanism (a) is mounted on the X-axis driving member.
9. The printing process of the three-dimensional ultrasonic array stentless cell printing device based on any of the claims 1-8, comprising the following steps,
s1: sterilizing the printing device, keeping the temperature constant and proper, injecting a culture solution into the polygonal resonant cavity, and injecting a cell suspension into the micro-nozzle;
s2: according to a pre-designed model, the piezoelectric transducer array works, and a planar array sound pressure micro-potential well is excited on the lowest layer of the polygonal resonant cavity as required;
s3: controlling the platform driving mechanism to move according to a pre-designed model to enable the spray head to reach a specific height required by printing of the polygonal resonant cavity model, and then controlling the conveying driving mechanism to move to enable the micro-nozzle to move to a specified sound pressure micro-potential well position;
s4: according to the requirement of a model, a cell conveying mechanism conveys a specific number of cells to the corresponding planar array sound pressure micro-potential well, the conveyed cells are captured in the sound pressure potential well under the action of the sound restoring force in the sound pressure potential well, the printing of the cells in a single potential well is completed, and the cell printing in all the sound potential wells in one layer of plane is completed in a sub-class push mode until the printing of the cells in all the sound potential wells in one layer of plane is completed;
s5: after the first layer is completely printed, the three-dimensional ultrasonic array micro-potential well generating system excites the array sound pressure micro-potential well array of the second layer, the polygonal culture platform is moved downwards to continue printing, the steps S2-S4 are repeated, and the printing of the cell three-dimensional system is realized by the analogy;
s6: and after printing, performing cell culture on the cell three-dimensional system at a proper temperature, and finally forming cell tissues through cell division and growth.
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| DE10018987A1 (en) * | 2000-04-17 | 2001-10-31 | Envision Technologies Gmbh | Device and method for producing three-dimensional objects |
| CN106399078B (en) * | 2015-07-29 | 2018-12-28 | 清华大学 | A kind of foreign cell 3 D-printing system and method |
| CN111826262B (en) * | 2019-04-18 | 2023-06-20 | 深圳先进技术研究院 | Biological 3D printing system and method |
| CN111254076B (en) * | 2020-01-19 | 2021-06-08 | 武汉大学 | Hexagonal surface wave acoustic tweezers chip for cell arrangement and assembly |
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